高压反相液相色谱法分析~3HCAMP与~3MCGMP

ANALYSIS OF ~3H CAMP AND ~3H CGMP BY HPLC

摘要: <正> CAMP(环磷酸腺苷)和CGMP(环磷酸鸟苷)属环核苷酸类,,在机体中的含量极低,约为每mg湿组织pg分子水平,用一般的分析方法不能测定。主要采用竞争性蛋白质结合分析法、放射免疫法和薄层色谱法进行分析。近年来,用高压液相色谱分离分析环核苷酸,定量方便、分析速度快、检测装置灵敏度高,具有其他分析方法所没有的优点。过去,对环核苷酸的分离都在离子交换柱上进行,最近,Krstulovic等采用反相色谱,在μBondapak C_(18)柱上进行分离分析,不需对样品进行预浓缩,使方法更为简便,而且柱寿命长。 ~3HCAMP是竞争性蛋白质结合分析法测定环磷酸腺苷,以及~3HCGMP是放射免疫法测定环磷酸鸟苷所必不可少的试剂,它们的放化纯将直接影响这两种分析方法的成败。本工作采用高压反相分配色谱法,在ODS术上,用甲醇-20 mmol/l KH_2PO_4作流动

Abstract: Using high pressure reversed phase liquid chromatography, ~3H CAMP and~3H CGMP were determined successfully. The column was HC-ODS-SIL-Xcolumn, and the moving phase was MeOH-20mmol/l KH_2PO_4 (10/90 or 3/97,V/V, pH=5.5). The analytical time was less than 10min. The measuring precisionwas 1.5% (UV) and 0.15% (radioactivity). The determination limit was 10~(-9)gand 10~2Bq with the detection efficiency of 62%.